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木醋杆菌纤维素合成的中间步骤:野生型和无纤维素突变体的全细胞及无细胞制剂研究

Intermediatry steps in Acetobacter xylinum cellulose synthesis: studies with whole cells and cell-free preparations of the wild type and a celluloseless mutant.

作者信息

Swissa M, Aloni Y, Weinhouse H, Benizman M

出版信息

J Bacteriol. 1980 Sep;143(3):1142-50. doi: 10.1128/jb.143.3.1142-1150.1980.

Abstract

Intermediatry steps in cellulose synthesis in Acetobacter xylinum were studied with resting cells and particulate-membranous preparations of the wild-type strain and of a celluloseless mutant. Exogenously supplied [1-14C]glucose was rapidly converted by resting cells of both types into glucose 6-phosphate, glucose 1-phosphate, and uridine glucose 5'-diphosphate (UDP)-glucose and incorporated into lipid-, water-, and alkali-soluble cellular fractions. The decrease in the level of labeled hexose-phosphates and UDP-glucose upon depletion of the exogenous substrate was accounted for by a continuous incorporation of [14C]glucose into cellulose in the wild type and into the above-mentioned cellular components in the mutant. [14C]glucose retained in the alkali- and water-soluble fractions of pulse-labeled wild-type cells was quantitatively chased into cellulose. Sonic extracts of both strains catalyzed the transfer of glucose from UDP-glucose into lipid-, water-, and alkali-soluble materials, as well as into an alkali-insoluble cellulosic beta-1,4-glucan. The results strongly support the sequence glucose leads to glucose 6-phosphate leads to glucose 1-phosphate leads to UDP-glucose leads to cellulose and indicate that lipid- and protein-linked cellodextrins may function as intermediates between UDP-glucose and cellulose in A. xylinum.

摘要

利用野生型菌株和无纤维素突变体的静息细胞以及颗粒膜制剂,研究了木醋杆菌中纤维素合成的中间步骤。两种类型的静息细胞都能将外源提供的[1-14C]葡萄糖迅速转化为6-磷酸葡萄糖、1-磷酸葡萄糖和尿苷葡萄糖5'-二磷酸(UDP)-葡萄糖,并掺入脂质、水和碱溶性细胞组分中。在外源底物耗尽后,标记的己糖磷酸和UDP-葡萄糖水平的下降,是由于野生型中[14C]葡萄糖持续掺入纤维素,而突变体中[14C]葡萄糖持续掺入上述细胞成分。脉冲标记的野生型细胞的碱溶性和水溶性组分中保留的[14C]葡萄糖被定量追踪到纤维素中。两种菌株的超声提取物都催化葡萄糖从UDP-葡萄糖转移到脂质、水和碱溶性物质中,以及转移到碱不溶性纤维素β-1,4-葡聚糖中。结果有力地支持了葡萄糖→6-磷酸葡萄糖→1-磷酸葡萄糖→UDP-葡萄糖→纤维素的序列,并表明脂质和蛋白质连接的纤维糊精可能在木醋杆菌中作为UDP-葡萄糖和纤维素之间的中间体发挥作用。

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